Acquisition of a Confocal Laser Scanning Microscope for Cell Biology and Biophysics

获得用于细胞生物学和生物物理学的共聚焦激光扫描显微镜项目概要伍兹霍尔海洋生物实验室（MBL）的Peter J.S. Smith博士获得了一笔拨款，用于获得具有多光子激发能力的共聚焦激光扫描显微镜和图像分析系统。该系统将被安置在MBL中央显微镜设施中，使其可供广泛的常驻研究人员使用，包括伍兹霍尔海洋研究所的科学家、访问和夏季研究人员，以及MBL提供的许多研究生和博士后水平的教育课程。调查小组已经制定了几个拟议的研究项目，涵盖基础生物学到生物医学科学。在MBL的居民社区中，这些包括：1。锥虫囊泡运输的研究。这是研究传染病的大型项目的一部分。Ca/CaM激酶II动力学在LTP中的作用。这是一个调查记忆基础的大型项目的一部分；3. 单细胞真核极端微生物的生理学。研究适应性进化和蛋白质表达/修饰的更大项目的一部分。病原体的多药耐药性。5. 有丝分裂纺锤体的结构动力学。正在进行的细胞动力学实时成像项目。神经元中的钙振荡。这是几个正在进行的研究细胞膜和细胞质中钙活性域的项目的组成部分。环境污染物对神经元发育的影响。这是MBL研究人员在毒理学领域研究的一个关键问题。红细胞铁运输的机制，形成了在MBL启动的一个新的线粒体程序的一部分。MBL的科学家们对细胞动力学的研究有着长期的兴趣，这必然需要对生命系统进行观察。要选择的显微镜可以最大限度地发挥这种能力。系统组件的选择是为了提供1)在可见光谱中不同荧光团的共聚焦荧光成像能力；2)多光子激发；3)发射指纹技术，用于同时记录具有紧密间隔发射光谱的多个荧光团；4)有足够范围的物镜，以进行活细胞的结构研究和生理实验；5)精确定位荧光标记结构；6)所研究活细胞的温度控制；7)软件充分利用共聚焦的数据采集能力；8)图像分析和显示软件。拨款中提出的研究活动反映了MBL社区对科学兴趣的多样性，以及在研究和教育方面的推广。新的显微镜将是一个宝贵的补充MBL设施的前沿研究。它对居民和游客的可用性（每年有超过200名季节性调查人员访问MBL进行研究）将确保高影响和使用所授予的赠款。

Acquisition of a Confocal Laser Scanning Microscope for Cell Biology and BiophysicsProject SummaryA grant has been awarded to Dr Peter J.S. Smith of the Marine Biological Laboratory (MBL), Woods Hole, for the acquisition of a confocal laser scanning microscope and image analysis system with multiphoton excitation capability. The system is to be housed in the MBL Central Microscopy Facility, making it available to a wide community of resident researchers, including scientists from the Woods Hole Oceanographic Institute, visiting and summer investigators, as well as by many of the graduate- and postdoctoral-level education courses offered at the MBLSeveral proposed research projects have already been formulated by the investigating team, spanning basic biology to the biomedical sciences. Amongst the resident community of the MBL these include:1. An examination of vesicle trafficking in trypanosomes. Part of a larger project dealing infectious diseases.2. The role of Ca/CaM kinase II dynamics in LTP. Part of a larger project investigating the foundations of memory; 3. The physiology of single-celled eukaryotic extremophiles. Part of a larger project to investigate adaptive evolution and protein expression/modification.4. Multidrug resistance in pathogens. 5. Architectural dynamics of the mitotic spindle. A continuing project on live imaging of cellular dynamics.6. Calcium oscillations in neurons. A component of several ongoing projects looking at calcium activity domains in the cell membrane and cytosol.7. The effects of environmental pollutants on neuronal development. A key issue being studied by MBL investigators in the field of toxicology.8. The mechanisms of iron trafficking in erythroid cells which forms part of a new mitochondrial program initiated at the MBL. Scientists at the MBL have had a long interest in the study of cellular dynamics, which by necessity requires observations on living systems. The microscope to be chosen maximizes this ability. System components were chosen to provide 1) capability for confocal fluorescent imaging of different fluorophores in the visible spectrum; 2) multiphoton excitation; 3) emission fingerprinting for simultaneous recording of multiple fluorophores with closely-spaced emission spectra; 4) a sufficient range of objective lenses to enable both structural studies and experiments on the physiology of living cells; 5) precisely localization fluorescent-labeled structures; 6) temperature control of live cells under study; 7) software to use fully the data acquisition capabilities of the confocal; and 8) software for image analysis and display. The research activities proposed in the grant reflect the diversity of scientific interest amongst the MBL community as well as outreach in research and education. The new microscope will be an invaluable addition the MBL facilities for cutting edge research. Its availability to residents and visitors alike (over 200 seasonal investigators visit the MBL for research purposes on an annual basis) will insure a high impact and use for the grant awarded.